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Tris Based Blocking Buffers

Product#: TrisBasedBlockingBuffers
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verified Phosphate-Free TBS / TBST Blocking · 0.1 µm + 0.04 µm Filtration

Tris Based Blocking Buffers - Category Selection Guide

Five phosphate-free blocking formulations on two Tris-buffered bases, ten catalog numbers in 500 ml and 1000 ml. This page is a category reference — pick the formulation your detection chemistry needs and click View for its product page; specifications, filtration architecture and selection guidance follow below.

Tris Based Blocking Buffers — Catalog · 10 Catalog Numbers Across 5 Product Pages
This page is a category reference, not a product page. Catalog numbers are grouped under the product page they order from; click View once per group.
Cat. No. Base Size
Bovine Serum Albumin (3%) with TBS Blocking Buffer TBS · 2 catalog numbers · 500 ml · 1000 ml
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DCP-BSATBS3X_500 ml TBS 500 ml
DCP-BSATBS3X_1000 ml TBS 1000 ml
Bovine Serum Albumin (3%) with TBST Blocking Buffer TBST · 2 catalog numbers · 500 ml · 1000 ml
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DCP-BSATBST3X_500 ml TBST 500 ml
DCP-BSATBST3X_1000 ml TBST 1000 ml
Casein (1%) with TBS Blocking Buffer TBS · 2 catalog numbers · 500 ml · 1000 ml
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DCP-CTBS1X_500 ml TBS 500 ml
DCP-CTBS1X_1000 ml TBS 1000 ml
Gelatin (1%) in TBS Blocking Buffer TBS · 2 catalog numbers · 500 ml · 1000 ml
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DCP-GTBS1X_500 ml TBS 500 ml
DCP-GTBS1X_1000 ml TBS 1000 ml
PVP in TBS Blocking Buffer TBS · 2 catalog numbers · 500 ml · 1000 ml
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DCP-PVPTBS1X_500 ml TBS 500 ml
DCP-PVPTBS1X_1000 ml TBS 1000 ml

Not sure which formulation? Compare the blocking agents · See applications by formulation · Read the FAQ

Family Snapshot

What the five formulations share, and where they part company

Tris-based blocking buffers provide an effective and versatile option for reducing non-specific binding in various protein detection techniques. Their phosphate-free nature makes them particularly suitable for phosphoprotein detection and alkaline phosphatase-based systems. The choice between different formulations depends on the specific requirements of the assay and the proteins being studied.

  • Five formulations: Bovine Serum Albumin (3%) with TBS, Bovine Serum Albumin (3%) with TBST, Casein (1%) with TBS, Gelatin (1%) in TBS, and PVP in TBS.
  • Two Tris-buffered bases: TBS and TBST, as published in the product names.
  • Phosphate-free: the source identifies the phosphate-free nature of these buffers as what makes them particularly suitable for phosphoprotein detection and alkaline phosphatase-based systems.
  • Appearance: Clear, Colorless Liquid.
  • Sterility: filtered 0.1 micron twice and 0.04 micron twice, in a sterile environment.
  • Fill sizes: every formulation is stocked in 500 ml and 1000 ml — ten catalog numbers in total.
  • Selection basis: the source states that the choice between formulations depends on the specific requirements of the assay and the proteins being studied.
PHOSPHATE-FREE 0.1 µm ×2 + 0.04 µm ×2 RUO
CATEGORY REFERENCE · TRIS BASED BLOCKING BUFFERS
The published specification, in the source’s own values
  • CategoryTris Based Blocking Buffers
  • Buffer bases stockedTBS, TBST
  • Formulations5
  • Catalog numbers10
  • Fill sizes500 ml, 1000 ml
  • AppearanceClear, Colorless Liquid
  • Filtration0.1 µm twice, 0.04 µm twice
  • Fill environmentsterile environment
  • Blocking agentsBSA 3%, Casein 1%, Gelatin 1%, PVP
  • Phosphate contentphosphate-free
Why The Blocking Agent Matters

Same base, different blocker, different blot

All five formulations sit on a Tris base, so the variable a buyer is actually choosing is the blocking agent. The published immunoassay literature is consistent on one point: that choice is not cosmetic. Each card below is cited to the reference list at the foot of the page.

blur_on

Non-specific binding is the target

The source describes these buffers as an effective and versatile option for reducing non-specific binding in various protein detection techniques. Blocking occupies the sites on the solid phase that the probe would otherwise adsorb to.

science

The Tris base is phosphate-free

The source attributes the fit of this family to that phosphate-free nature, which it says makes these buffers particularly suitable for phosphoprotein detection and alkaline phosphatase-based systems.

compare_arrows

The blocker changes what you detect

A direct comparison of serum albumin, non-fat dry milk and Tween 20 on nitrocellulose found quantitative and qualitative differences in the antigens detected, and recommended comparing several blocking and washing methods rather than assuming one.[2]

bar_chart

Blocking efficiency differs by agent

Across a million-fold concentration range on polystyrene plates, casein and instantized milk suppressed non-specific binding by over 90 % at far lower concentrations than most of eight other proteins; enzymatically hydrolysed porcine-skin gelatin was the least effective protein tested.[3]

warning

Casein is itself a phosphoprotein

Casein is the primary blocking protein of non-fat dry milk and is a phosphoprotein, which is why casein-based blocking is reported as unsuitable for phospho-specific antibodies such as anti-phosphoserine and anti-phosphothreonine.[7]

tune

A protein-free route exists

Polyvinylpyrrolidone was described as a blocking agent for immunochemical work precisely because it is not a protein[4]; it is also the long-standing blocking component of Denhardt’s hybridisation solution.[5]

The one decision this page exists to support

The source says the choice between formulations depends on the specific requirements of the assay and the proteins being studied — and the literature says where that bites hardest: a casein blocker and a phospho-specific antibody are a documented mismatch[7], while a purified single protein such as BSA carries its own caveat that preparations differ and need a blocking control.[6]

4
membrane passes stated per lot — 0.1 µm twice, then 0.04 µm twice
10
catalog numbers across 5 product pages, in 500 ml and 1000 ml
Filtration Architecture

Four membrane passes, ending at 0.04 micron

The source states one sterility line for the whole family: filtered 0.1 micron twice and 0.04 micron in a sterile environment. Under the canonical house wording that is a four-stage architecture, and it is the same for every catalog number on this page.

  1. 1

    0.1 µm Pre-filtration I

    First pass through a 0.1 micron membrane, removing the bulk particulate and microbial load before the finer stages.

  2. 2

    0.04 µm Pre-filtration II

    Sub-0.1 micron pass, taking out the fine fraction that a 0.1 micron membrane passes.

  3. 3

    0.1 µm Sterile-filtration I

    Second 0.1 micron pass, performed in the sterile environment the source specifies for the fill.

  4. 4

    0.04 µm Sterile-filtration II — Final Polish

    Final 0.04 micron polish before fill. The delivered solution is a Clear, Colorless Liquid.

Quadruple-stage filtration diagram for Diagnocine Tris Based Blocking Buffers: 0.1 micron membrane pre-filtration twice and 0.04 micron membrane polishing twice, performed in a sterile environment, for TBS and TBST blocking buffers used in protein detection and phosphoprotein workflows
Figure 1. The four-stage filtration architecture stated for every Tris Based Blocking Buffer catalog number on this page: 0.1 µm twice, then 0.04 µm twice, in a sterile environment. © Diagnocine® — Tris Based Blocking Buffers
On the wording. The source line reads “Filtered 0.1 micron Twice and 0.04 micron once in a sterile environment”. House rule 24 canonicalizes that exact pattern to 0.04 micron twice; both phrasings resolve to the same four-stage architecture and the same figure. Filtration method and pass count can change per order — confirm the architecture on the Certificate of Analysis for the lot you receive, or ask support@diagnocine.com.
Blocking Agent Reference

The five stocked agents, and what the literature records about each

The source does not publish a comparison table for this family, so the table below pairs each stocked formulation with the published characterisation of its blocking agent. Every entry is cited; nothing in the “Formulation” column has been altered from the catalog.

Formulation (as stocked) Agent class What the published literature records Watch-out
Bovine Serum Albumin (3%) with TBS Blocking Buffer Single purified protein Serum albumin is one of the standard blockers benchmarked against milk, casein and gelatin on plates and on nitrocellulose.[2,3] BSA preparations are not interchangeable: non-specific binding to BSA has been traced to the preparation itself, so a blocking-agent control is advised.[6]
Bovine Serum Albumin (3%) with TBST Blocking Buffer Single purified protein on a Tween-containing base The non-ionic detergent Tween 20 was itself introduced as a means of blocking unoccupied protein-binding sites on nitrocellulose, in place of a protein.[1] Tween 20 used alone as the blocking agent has been reported to give artefactual results, so it is generally used alongside a protein blocker rather than instead of one.[9]
Casein (1%) with TBS Blocking Buffer Milk phosphoprotein Casein and instantized milk were the most effective proteins tested for suppressing non-specific binding of a peroxidase conjugate, blocking by over 90 % at far lower concentrations than most alternatives.[3] Casein is a phosphoprotein; casein-based blocking is reported as unsuitable for phospho-specific antibodies.[7]
Gelatin (1%) in TBS Blocking Buffer Denatured collagen Blocking performance depends strongly on the gelatin source: fish-skin gelatin blocked far better than enzymatically hydrolysed porcine-skin gelatin, which was the weakest protein in the series and was almost useless as a pre-treatment agent.[3] The source does not state the gelatin origin for this catalog item. Confirm the source before assuming a performance figure from the literature applies.
PVP in TBS Blocking Buffer Synthetic polymer, protein-free Polyvinylpyrrolidone was characterised as a blocking agent for immunochemical studies[4], and is the classic blocking polymer of Denhardt’s membrane-hybridisation solution.[5] The source does not state a PVP concentration for this catalog item, while the other four formulations carry one.
One clarification beyond the source (delete if unwanted). Tris is a comparatively temperature-sensitive buffer: a Tris-based solution adjusted to a given pH at room temperature does not sit at that same pH at 4 °C or 37 °C, which is worth knowing when blocking, incubation and washing steps run at different temperatures.[8] This point is not in the Diagnocine source description; it is added for buyers moving from a phosphate base to a Tris base.
Applications by Formulation

Which formulation for which detection chemistry

The source assigns applications to the category as a whole, not to individual formulations. Each tab therefore states the source’s own category-level use first, then the published characterisation of that specific blocking agent, cited. Select a formulation to see both.

Bovine Serum Albumin (3%) · TBS · phosphate-free
  • Reducing non-specific binding in various protein detection techniques (source).
  • Phosphoprotein detection and alkaline phosphatase-based systems, which the source attributes to the phosphate-free Tris base.
  • Workflows where a single purified protein is preferred over a milk-derived blocker — casein-based blocking is reported as unsuitable for phospho-specific antibodies.[7]
  • Run a blocking-agent control: non-specific binding has been traced to the BSA preparation itself rather than to the assay.[6]
Bovine Serum Albumin (3%) · TBST · Tween-containing base
  • Reducing non-specific binding in various protein detection techniques (source).
  • Protocols that keep a non-ionic detergent in the blocking step; Tween 20 was introduced as a way of blocking unoccupied protein-binding sites on nitrocellulose in place of a protein.[1]
  • Phosphoprotein detection and alkaline phosphatase-based systems, on the same phosphate-free Tris base as the TBS version (source).
  • Note that Tween 20 used alone as the blocking agent has been reported to produce artefactual results.[9]
Casein (1%) · TBS · strongest blocker in the series
  • Reducing non-specific binding in various protein detection techniques (source).
  • General protein detection where blocking strength is the priority: casein blocked non-specific binding by over 90 % at far lower concentrations than most of eight other proteins tested.[3]
  • Not the formulation to reach for with phospho-specific antibodies — casein is itself a phosphoprotein.[7]
Gelatin (1%) · TBS · denatured collagen
  • Reducing non-specific binding in various protein detection techniques (source).
  • An alternative when albumin- or milk-derived blockers cross-react with the probe; gelatin sits in the same benchmarked series as albumin, casein and milk.[3]
  • Published performance is source-dependent: fish-skin gelatin blocked far better than enzymatically hydrolysed porcine-skin gelatin.[3] The Diagnocine source does not state which gelatin this item uses — confirm before transferring a literature figure to it.
PVP · TBS · protein-free
  • Reducing non-specific binding in various protein detection techniques (source).
  • Workflows in which every protein blocker is a potential cross-reactant: polyvinylpyrrolidone was characterised as a blocking agent for immunochemical studies precisely because it is not a protein.[4]
  • Membrane-hybridisation lineage: PVP is the classic blocking polymer of Denhardt’s solution.[5]
  • Phosphoprotein detection and alkaline phosphatase-based systems, on the phosphate-free Tris base (source).
Product Comparison

The five formulations side by side

Every value in this table is reproduced from the catalog exactly as published.

Product Blocking agent Base Catalog numbers Sizes
Bovine Serum Albumin (3%) with TBS Blocking Buffer Bovine Serum Albumin (3%) TBS DCP-BSATBS3X_500 ml
DCP-BSATBS3X_1000 ml		 500 ml, 1000 ml
Bovine Serum Albumin (3%) with TBST Blocking Buffer Bovine Serum Albumin (3%) TBST DCP-BSATBST3X_500 ml
DCP-BSATBST3X_1000 ml		 500 ml, 1000 ml
Casein (1%) with TBS Blocking Buffer Casein (1%) TBS DCP-CTBS1X_500 ml
DCP-CTBS1X_1000 ml		 500 ml, 1000 ml
Gelatin (1%) in TBS Blocking Buffer Gelatin (1%) TBS DCP-GTBS1X_500 ml
DCP-GTBS1X_1000 ml		 500 ml, 1000 ml
PVP in TBS Blocking Buffer PVP (concentration not stated in source) TBS DCP-PVPTBS1X_500 ml
DCP-PVPTBS1X_1000 ml		 500 ml, 1000 ml
FAQ

Frequently asked questions

The questions that come up when a purchasing decision meets a detection chemistry.

The blocking agent is carried on a Tris-buffered saline base rather than a phosphate base. According to the source, Tris-based blocking buffers are an effective and versatile option for reducing non-specific binding in various protein detection techniques, and their phosphate-free nature makes them particularly suitable for phosphoprotein detection and alkaline phosphatase-based systems.
The source states only that the choice depends on the specific requirements of the assay and the proteins being studied. The published literature narrows it in one direction: casein is a phosphoprotein and is the primary blocking protein of non-fat dry milk, which is why casein-based blocking is reported as unsuitable for phospho-specific antibodies such as anti-phosphoserine and anti-phosphothreonine.[7] That points away from the Casein (1%) formulation and toward one of the others on this page.
They are published as two separate catalog items with the same blocking agent, Bovine Serum Albumin (3%), on two different bases — TBS and TBST. TBST carries the non-ionic detergent Tween 20, which was itself introduced as a way of blocking unoccupied protein-binding sites on nitrocellulose in place of a protein.[1] Note that Tween 20 used alone as the blocking agent has been reported to produce artefactual results.[9]
The source gives one sterility line for the whole family: filtered 0.1 micron twice and 0.04 micron in a sterile environment, canonicalized here to 0.1 micron twice and 0.04 micron twice — a four-stage architecture ending in a sub-0.1 micron polish. The delivered solution is described as a Clear, Colorless Liquid. Filtration method and pass count can change per order, so confirm against the Certificate of Analysis for the lot you receive.
The catalog item is published as Casein (1%) with TBS Blocking Buffer, not as milk. Casein is the primary blocking protein of non-fat dry milk[7], and in a plate study casein and instantized milk were the two most effective proteins tested, suppressing non-specific binding by over 90 % at far lower concentrations than most of eight other proteins.[3] The phospho caveat that applies to milk applies here for the same reason: casein is a phosphoprotein.[7]
When any protein in the blocking solution is a potential cross-reactant for the probe. Polyvinylpyrrolidone was characterised as a blocking agent for immunochemical studies precisely because it is not a protein[4], and it is the long-standing blocking polymer of Denhardt’s membrane-hybridisation solution.[5] The source does not state a PVP concentration for this catalog item, so ask before assuming one.
Every formulation is stocked in 500 ml and 1000 ml, giving ten catalog numbers across five product pages: DCP-BSATBS3X, DCP-BSATBST3X, DCP-CTBS1X, DCP-GTBS1X and DCP-PVPTBS1X, each with a _500 ml and a _1000 ml suffix. The catalog table at the top of this page groups the catalog numbers under the product page they order from.
Key References

Supporting literature on blocking agents

The Diagnocine source description carries no citation list of its own. The peer-reviewed sources below are the ones cited in the sections above; each was checked against its published record before it was listed.

  1. Batteiger B, Newhall WJ 5th, Jones RB (1982). The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods 55(3):297–307. doi:10.1016/0022-1759(82)90089-8
  2. Spinola SM, Cannon JG (1985). Different blocking agents cause variation in the immunologic detection of proteins transferred to nitrocellulose membranes. J Immunol Methods 81(1):161–165. doi:10.1016/0022-1759(85)90132-2
  3. Vogt RF Jr, Phillips DL, Henderson LO, Whitfield W, Spierto FW (1987). Quantitative differences among various proteins as blocking agents for ELISA microtiter plates. J Immunol Methods 101(1):43–50. doi:10.1016/0022-1759(87)90214-6
  4. Haycock JW (1993). Polyvinylpyrrolidone as a blocking agent in immunochemical studies. Anal Biochem 208(2):397–399. doi:10.1006/abio.1993.1068
  5. Denhardt DT (1966). A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun 23(5):641–646. doi:10.1016/0006-291X(66)90447-5
  6. Xiao Y, Isaacs SN (2012). Enzyme-linked immunosorbent assay (ELISA) and blocking with bovine serum albumin (BSA) — not all BSAs are alike. J Immunol Methods 384(1–2):148–151. doi:10.1016/j.jim.2012.06.009
  7. Galva C, Gatto C, Milanick M (2012). Soymilk: an effective and inexpensive blocking agent for immunoblotting. Anal Biochem 426(1):22–23. doi:10.1016/j.ab.2012.03.028 — source for the statement that casein is the primary blocking protein of non-fat dry milk and is a phosphoprotein, making it unsuitable for phospho-specific antibodies.
  8. Good NE, Winget GD, Winter W, Connolly TN, Izawa S, Singh RMM (1966). Hydrogen ion buffers for biological research. Biochemistry 5(2):467–477. doi:10.1021/bi00866a011
  9. Bird CR, Gearing AJH, Thorpe R (1988). The use of Tween 20 alone as a blocking agent for immunoblotting can cause artefactual results. J Immunol Methods 106(2):175–179.
Formulation selection support. For help matching a blocking formulation to a specific protocol, for the gelatin source or PVP concentration behind a catalog item, or for documentation requests, contact support@diagnocine.com. Ready to order? Back to the Tris Based Blocking Buffers catalog.

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